Compounds affecting pigment production and methods for treatment of bacterial diseases

ABSTRACT

The present methods include administering to a subject an effective amount of one or more compounds of Formula (II). In one embodiment, said microbial infections are bacterial infections. More specifically, said bacterial infections are staphylococcal infections.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority from a U.S. provisional patentapplication Ser. No. 62/535,540 filed Jul. 21, 2017, and the disclosureof which is incorporated by reference in its entirety.

TECHNICAL FIELD

The present invention relates to compounds and derivatives thereof,composition comprising said compounds and/or derivatives, and methodsfor treating microbial infections and/or related diseases or conditions.More specifically, the present compounds, derivatives, compositioncomprising thereof, and methods are for bacterial infections.

BACKGROUND OF THE INVENTION

Staphylococcus aureus is a major human pathogen in communities andhospitals, causing a variety of infections that ranges from harmlessinfections to life threatening conditions [18]. With the wide-spreaddissemination of methicillin-resistant S. aureus (MRSA) in hospitals andin communities, treating S. aureus associated infections has becomeincreasingly difficult [19]. Staphyloxanthin has been proven to be animportant factor in promoting bacterial invasion [1]. Five genes,crtOPQMN, located in an operon are responsible for the biosynthesis ofthe pigment. The transcription of the operon is driven by aσ^(B)-dependent promoter upstream of crtO, and ends with a terminatordownstream of crtN [2]. The pigments that endow S. aureus with a goldencolor also make it resistant to attack from reactive oxygen species(ROS) and neutrophils [3]. Pigmented bacteria have increased resistanceto the host's immune defenses [4].

In a mouse subcutaneous model of infection, animals infected with awild-type strain of S. aureus had higher bacterial loads and largervisible lesions than those infected with non-pigmented bacteria [4]. Thereduced virulence of bacterial strains with defective carotenoidsynthesis was also shown in a mouse systemic S. aureus infection model[3]. In vitro and in vivo data suggest that blocking pigment synthesismay reduce pathogenicity.

Dehydrosqualene synthase (CrtM) catalyses the first step of thebiosynthetic pathway, was shown to be a target for anti-infectivetherapy based on virulence factor neutralization. Diphenylamine wasfound to be an inhibitor of 4,4-diapophytoene desaturase (CrtN) at highmicromolar level [5]. Another potential inhibitor of CrtN, naftifine, aFDA approved antifungal compound was shown to reduce bacterial load indifferent mice infection models [6]. However, there remains a need fornew compounds and methods of treatment for staphylococcal infections.

SUMMARY OF THE INVENTION

Provided herein are compounds and methods for prevention and/ortreatment of microbial infections and/or related disease or conditions.In a first aspect, the present invention provides compounds and/or theirderivatives which can be represented by Formula (II):

wherein R1 is selected from:

or

-   any four-, five-, six-, seven-, eight-, nine-, ten-, eleven-, or    twelve-membered heterocyclyl, cycloalkenyl, or cycloalkyl,-   where R3 and R4 can be independently or jointly selected from the    group: H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl, such as methyl, ethyl,    n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;    (C₂₋₄)alkenyl, such as ethenyl, propenyl, butenyl, where the double    bond can be located at any position in the alkenyl carbon chain, and    including any alkenyl conformational isomers thereof; alkynyl;    aralkyl; alkaryl; halogenated alkyl; heteroalkyl; aryl;    heterocyclyl; cycloalkyl; cycloalkenyl; cycloalkynyl; hydroxyalkyl;    aminoalkyl; amino; alkylamino; arylamino; dialkylamino;    alkylarylamino; diarylamino; acylamino; hydroxyl; thiol; thioalkyl;    alkoxy; alkylthio; alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro;    carbamoyl; trifluoromethyl; phenoxy; benzyloxy; phosphonic acid;    phosphate ester; sulfonic acid (—SO₃H); sulfonate ester;    sulfonamide; alkaryl; arylalkyl; carbamate; amino; alkylamino;    arylamino; dialkylamino; alkylarylamino; diarylamino; alkylthio;    heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone (═O);    ether (—OR10); and ester (—COOR11 and —OC(═O)R11);-   or R3 and R4 can be bonded together to form a four-, five-, or    six-membered heterocyclyl, cycloalkenyl, or cycloalkyl;-   R5 can be selected from the group: H; F; Cl; Br; I; OH; CN;    (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl,    sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl, such as ethenyl,    propenyl, butenyl, where the double bond can be located at any    position in the alkenyl carbon chain, and including any alkenyl    conformational isomers thereof; alkynyl; aralkyl; alkaryl;    halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;    cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino;    alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;    acylamino; hydroxyl; thiol; thioalkyl; alkoxy; alkylthio;    alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro; carbamoyl;    trifluoromethyl; phenoxy; benzyloxy; phosphonic acid; phosphate    ester; sulfonic acid (—SO₃H); sulfonate ester; sulfonamide; alkaryl;    arylalkyl; carbamate; amino; alkylamino; arylamino; dialkylamino;    alkylarylamino; diarylamino; alkylthio; heteroalkyl;    alkyltriphenylphosphonium; heterocyclyl; ketone (═O); ether (—OR10);    and ester (—COOR11 and —OC(═O)R11); and-   where R10 and R11 can be independently or jointly selected from the    group consisting of: a (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers; and alkynyl;-   X is selected from N or C,-   A is single bond or double bond;-   Q is selected from N or C,-   M is selected from O or C, and-   wherein R2 is selected from:

where R6 and R7 can be independently or jointly selected from O orabsent;

-   R8 and R9 can be independently or jointly selected from H; F; Cl;    Br; I; OH; CN; (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers thereof; alkynyl; aralkyl;    alkaryl; halogenated alkyl; heteroalkyl; aryl; heterocyclyl;    cycloalkyl; cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl;    amino; alkylamino; arylamino; dialkylamino; alkylarylamino;    diarylamino; acylamino; hydroxyl; thiol; thioalkyl; alkoxy;    alkylthio; alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro;    carbamoyl; trifluoromethyl; phenoxy; benzyloxy; phosphonic acid;    phosphate ester; sulfonic acid (—SO₃H); sulfonate ester;    sulfonamide; alkaryl; arylalkyl; carbamate; amino; alkylamino;    arylamino; dialkylamino; alkylarylamino; diarylamino; alkylthio;    heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone (═O);    ether (—OR10); and ester (—COOR11 and —OC(═O)R11),-   or R8 and R9 can be bonded together to form a four-, five-, or    six-membered heterocyclyl, cycloalkenyl, or cycloalkyl. and-   where R10 and R11 can be independently or jointly selected from the    group consisting of: a (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers; and alkynyl, and-   Z is selected from C or S.

In one embodiment, the present compounds and/or the derivatives thereofcan be an anti-virulent agent for bacteria.

In another embodiment, the present compounds and/or the derivativesthereof are effective in reducing virulence of bacteria.

In other embodiment, the bacteria that the present compounds and/or thederivatives thereof are effective in reducing their virulence compriseStaphylococci sp.

In yet another embodiment, the bacteria that the present compoundsand/or the derivatives thereof are effective in reducing their virulencecomprise Staphylococcus aureus (S. aureus).

In still another embodiment, the bacteria that the present compoundsand/or the derivatives thereof are effective in reducing their virulencecomprise methicillin-resistant S. aureus (MRSA).

In other embodiment, said reducing the virulence of bacteria by thecompounds and/or derivatives thereof comprises inhibiting biosynthesisof staphyloxanthin in said bacteria and/or inhibiting or reducingproduction of pigments that are resistant to the bacterial host's immunedefenses.

A composition for preventing and/or treating the microbial infectionsand/or related diseases or conditions comprising an effective amount ofthe compounds and/or the derivatives thereof in the first aspect is alsoprovided herein.

In one embodiment, said microbial infections are bacterial infections.

In another embodiment, said microbial infections comprise staphylococcalinfections.

In other embodiment, the composition further comprises apharmaceutically acceptable carrier, salt, ester, expicient, vehicle,prodrug, solvent, and diluent, or any combination thereof.

In a second aspect, the present invention provides methods forpreventing and/or treating the microbial infections and/or relateddiseases or conditions including administering to a subject acomposition comprising an effective amount of one or more compounds ofFormula (II):

wherein R1 is selected from:

or

-   any four-, five-, six-, seven-, eight-, nine-, ten-, eleven-, or    twelve-membered heterocyclyl, cycloalkenyl, or cycloalkyl,-   where R3 and R4 can be independently or jointly selected from the    group: H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl, such as methyl, ethyl,    n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;    (C₂₋₄)alkenyl, such as ethenyl, propenyl, butenyl, where the double    bond can be located at any position in the alkenyl carbon chain, and    including any alkenyl conformational isomers thereof; alkynyl;    aralkyl; alkaryl; halogenated alkyl; heteroalkyl; aryl;    heterocyclyl; cycloalkyl; cycloalkenyl; cycloalkynyl; hydroxyalkyl;    aminoalkyl; amino; alkylamino; arylamino; dialkylamino;    alkylarylamino; diarylamino; acylamino; hydroxyl; thiol; thioalkyl;    alkoxy; alkylthio; alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro;    carbamoyl; trifluoromethyl; phenoxy; benzyloxy; phosphonic acid;    phosphate ester; sulfonic acid (—SO₃H); sulfonate ester;    sulfonamide; alkaryl; arylalkyl; carbamate; amino; alkylamino;    arylamino; dialkylamino; alkylarylamino; diarylamino; alkylthio;    heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone (═O);    ether (—OR10); and ester (—COOR11 and —OC(═O)R11);-   or R3 and R4 can be bonded together to form a four-, five-, or    six-membered heterocyclyl, cycloalkenyl, or cycloalkyl;-   R5 can be selected from the group: H; F; Cl; Br; I; OH; CN;    (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl,    sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl, such as ethenyl,    propenyl, butenyl, where the double bond can be located at any    position in the alkenyl carbon chain, and including any alkenyl    conformational isomers thereof; alkynyl; aralkyl; alkaryl;    halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;    cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino;    alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;    acylamino; hydroxyl; thiol; thioalkyl; alkoxy; alkylthio;    alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro; carbamoyl;    trifluoromethyl; phenoxy; benzyloxy; phosphonic acid; phosphate    ester; sulfonic acid (—SO₃H); sulfonate ester; sulfonamide; alkaryl;    arylalkyl; carbamate; amino; alkylamino; arylamino; dialkylamino;    alkylarylamino; diarylamino; alkylthio; heteroalkyl;    alkyltriphenylphosphonium; heterocyclyl; ketone (═O); ether (—OR10);    and ester (—COOR11 and —OC(═O)R11); and-   where R10 and R11 can be independently or jointly selected from the    group consisting of: a (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers; and alkynyl;-   X is selected from N or C,-   A is single bond or double bond;-   Q is selected from N or C,-   M is selected from O or C, and-   wherein R2 is selected from:

where R6 and R7 can be independently or jointly selected from O orabsent;

-   R8 and R9 can be independently or jointly selected from H; F; Cl;    Br; I; OH; CN; (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers thereof; alkynyl; aralkyl;    alkaryl; halogenated alkyl; heteroalkyl; aryl; heterocyclyl;    cycloalkyl; cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl;    amino; alkylamino; arylamino; dialkylamino; alkylarylamino;    diarylamino; acylamino; hydroxyl; thiol; thioalkyl; alkoxy;    alkylthio; alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro;    carbamoyl; trifluoromethyl; phenoxy; benzyloxy; phosphonic acid;    phosphate ester; sulfonic acid (—SO₃H); sulfonate ester;    sulfonamide; alkaryl; arylalkyl; carbamate; amino; alkylamino;    arylamino; dialkylamino; alkylarylamino; diarylamino; alkylthio;    heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone (═O);    ether (—OR10); and ester (—COOR11 and —OC(═O)R11),-   or R8 and R9 can be bonded together to form a four-, five-, or    six-membered heterocyclyl, cycloalkenyl, or cycloalkyl. and-   where R10 and R11 can be independently or jointly selected from the    group consisting of: a (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers; and alkynyl, and-   Z is selected from C or S.

In one embodiment, the microbial infections are bacterial infection.

In another embodiment, the microbial infections comprise Staphylococcalinfections.

In other embodiment, the microbial infections and/or related diseases orconditions are caused by Staphylococci sp.

In yet another embodiment, the Staphylococci sp. comprise Staphylococcusaureus (S. aureus).

In still another embodiment, S. aureus comprise methicillin-resistant S.aureus (MRSA).

In other embodiment, the microbial infections and/or related diseases orconditions comprise infections of the skin and soft tissue, bone andjoint, surgical wound, indwelling devices, lung and heart valves.

In certain embodiments, the present method further comprises reducingvirulence of bacteria causing the microbial infections and/or relateddisease or conditions.

In some other embodiments, the present method further comprisesinhibiting biosynthesis of staphyloxanthin in said bacteria and/orinhibiting or reducing production of pigments that are resistant to thebacterial host's immune defenses.

In another embodiment, said subject or bacterial host is a mammal.

In other embodiment, said subject or bacterial host is human.

BRIEF DESCRIPTION OF THE DRAWINGS

In the following detailed description, reference is made to theaccompanying figures, depicting exemplary, non-limiting andnon-exhaustive embodiments of the invention. So that the manner in whichthe above recited features of the present invention can be understood indetail, a more particular description of the invention, brieflysummarized above, can be had by reference to the embodiments, some ofwhich are illustrated in the appended figures. It should be noted,however, that the figures illustrate only typical embodiments of thisinvention and are therefore not to be considered limiting of its scope,for the invention can admit to other equally effective embodiments.

FIGS. 1A-1D show the in vitro pigment inhibition by compound NP16: FIG.1A shows the inhibition of wild-type (WT) S. aureus pigmentation usingincreasing concentrations of NP16; FIG. 1B shows the pigment inhibitionby NP16; the IC₅₀ for pigment formation is ˜300 nM; FIG. 1C depicts thechemical structure of compound NP16; FIG. 1D shows the growth curve ofS. aureus COL in the presence of different concentrations of NP16. Alldata represent mean values±SD.

FIGS. 2A-2D show that NP16 treatment leads to increased sensitivity tooxidation and neutrophil killing: FIG. 2A depicts the cytotoxic activityof compound NP16 on MDCK cells; FIG. 2B shows the increasedsusceptibility of the NP16-treated S. aureus COL strain to killing byhydrogen peroxide; FIG. 2C shows the increased susceptibility of theNP16-treated S. aureus COL to killing by neutrophils; FIG. 2D is the UVspectrum of carotenoids extracted from different strains, with orwithout NP16 treatment. All data represent mean values±SD (***P<0.001;****P<0.0001). P values were determined using GraphPad Prism using anunpaired parametric t test with Welch's correction.

FIGS. 3A-3F show the in vivo effect of CrtN and its inhibition by NP16.FIGS. 3A and 3B show the bacteria recovered from the livers and spleens,respectively, of mice infected with the wild-type COL or COL-ΔcrtNstrains; FIGS. 3C and 3D show the bacteria recovered from the livers andspleens, respectively, of mice infected with the COL strain, with orwithout compound NP16 treatment; FIG. 3E shows the bacteria recoveredfrom the kidneys of mice infected with clinical isolate strain AE052 orAE052-ΔcrtN; FIG. 3F shows the bacteria recovered from the kidneys ofmice infected with strain AE052, with or without compound NP16treatment. All data represent mean values±SEM (*P<0.05; **P<0.01;***P<0.001). P values were determined using GraphPad Prism using anunpaired parametric t test with Welch's correction.

FIG. 4 shows in vivo efficacy of staphyloxanthin inhibitors fromselected NP-16 analogues.

DETAILED DESCRIPTION OF THE INVENTION

Following an established screening method for finding agents that reduceStaphyloccous aureus pigmentation [7], it is identified that the presentcompounds, termed NP16 and its derivatives, have block pigmentproduction in S. aureus by targeting the 4,4-diapophytoene desaturase(CrtN). CrtN is proposed as a novel target for anti-virulence treatmentsin S. aureus. S. aureus staphyloxanthin contributes substantially topathogenesis by interfering with host immune clearance mechanisms, buthas little impact on ex vivo survival of the bacteria. Without wantingto be bound by theory, it is provided that agents blockingstaphyloxanthin production may discourage the establishment andmaintenance of bacterial infection without exerting selective pressurefor antimicrobial resistance.

NP16 and its derivatives can be represented by Formula (II):

wherein R1 is selected from:

or

-   any four-, five-, six-, seven-, eight-, nine-, ten-, eleven-, or    twelve-membered heterocyclyl, cycloalkenyl, or cycloalkyl,-   where R3 and R4 can independently or jointly be selected from the    group: H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl, such as methyl, ethyl,    n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;    (C₂₋₄)alkenyl, such as ethenyl, propenyl, butenyl, where the double    bond can be located at any position in the alkenyl carbon chain, and    including any alkenyl conformational isomers thereof; alkynyl;    aralkyl; alkaryl; halogenated alkyl; heteroalkyl; aryl;    heterocyclyl; cycloalkyl; cycloalkenyl; cycloalkynyl; hydroxyalkyl;    aminoalkyl; amino; alkylamino; arylamino; dialkylamino;    alkylarylamino; diarylamino; acylamino; hydroxyl; thiol; thioalkyl;    alkoxy; alkylthio; alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro;    carbamoyl; trifluoromethyl; phenoxy; benzyloxy; phosphonic acid;    phosphate ester; sulfonic acid (—SO₃H); sulfonate ester;    sulfonamide; alkaryl; arylalkyl; carbamate; amino; alkylamino;    arylamino; dialkylamino; alkylarylamino; diarylamino; alkylthio;    heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone (═O);    ether (—OR10); and ester (—COOR11 and —OC(═O)R11);-   or R3 and R4 can be bonded together to form a four-, five-, or    six-membered heterocyclyl, cycloalkenyl, or cycloalkyl;-   R5 can be selected from the group: H; F; Cl; Br; I; OH; CN;    (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl,    sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl, such as ethenyl,    propenyl, butenyl, where the double bond can be located at any    position in the alkenyl carbon chain, and including any alkenyl    conformational isomers thereof; alkynyl; aralkyl; alkaryl;    halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;    cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino;    alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;    acylamino; hydroxyl; thiol; thioalkyl; alkoxy; alkylthio;    alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro; carbamoyl;    trifluoromethyl; phenoxy; benzyloxy; phosphonic acid; phosphate    ester; sulfonic acid (—SO₃H); sulfonate ester; sulfonamide; alkaryl;    arylalkyl; carbamate; amino; alkylamino; arylamino; dialkylamino;    alkylarylamino; diarylamino; alkylthio; heteroalkyl;    alkyltriphenylphosphonium; heterocyclyl; ketone (═O); ether (—OR10);    and ester (—COOR11 and —OC(═O)R11); and-   where R10 and R11 can be independently or jointly selected from the    group consisting of: a (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers; and alkynyl;-   X is selected from N or C,-   A is single bond or double bond;-   Q is selected from N or C,-   M is selected from O or C, and-   wherein R2 is selected from:

where R6 and R7 can be independently or jointly selected from O orabsent;

-   R8 and R9 can be independently or jointly selected from H; F; Cl;    Br; I; OH; CN; (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers thereof; alkynyl; aralkyl;    alkaryl; halogenated alkyl; heteroalkyl; aryl; heterocyclyl;    cycloalkyl; cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl;    amino; alkylamino; arylamino; dialkylamino; alkylarylamino;    diarylamino; acylamino; hydroxyl; thiol; thioalkyl; alkoxy;    alkylthio; alkoxyalkyl; aryloxy; arylalkoxy; acyloxy; nitro;    carbamoyl; trifluoromethyl; phenoxy; benzyloxy; phosphonic acid;    phosphate ester; sulfonic acid (—SO₃H); sulfonate ester;    sulfonamide; alkaryl; arylalkyl; carbamate; amino; alkylamino;    arylamino; dialkylamino; alkylarylamino; diarylamino; alkylthio;    heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone (═O);    ether (—OR10); and ester (—COOR11 and —OC(═O)R11), or R8 and R9 can    be bonded together to form a four-, five-, or six-membered    heterocyclyl, cycloalkenyl, or cycloalkyl. and-   where R10 and R11 can be independently or jointly selected from the    group consisting of: a (C₁₋₄)alkyl, such as methyl, ethyl, n-propyl,    isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl,    such as ethenyl, propenyl, butenyl, where the double bond can be    located at any position in the alkenyl carbon chain, and including    any alkenyl conformational isomers; and alkynyl, and-   Z is selected from C or S.

The compounds of Formula (II) can include, but are not limited to, thosecompounds listed in Table 1.

TABLE 1 Compounds Blocking Staphyloxanthin Production Sample nameChemical Name Structure NP16 3-phenyl-N-[4-(1-pyrrolidinylsulfonyl)phenyl]acryl- amide

NP16-XL- 010 3-phenyl-N-[4-(1-piperidine-1- sulfonyl)phenyl]acrylamide

NP16-XL- 011 3-(4-acetoxylphenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 012 3-(5-acetoxylphenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 013 3-(6-acetoxylphenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 014 3-(4-bromophenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 015 3-(5-bromophenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 016 3-(6-bromophenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 017 3-(4-methylphenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 018 3-(6-methylphenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 019 3-phenyl-N-[4-(1-indole-1- sulfonyl)phenyl]acrylamide

NP16-XL- 020 3-(4-bromophenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 021 3-(5-bromophenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 022 3-phenyl-N-[4-(1- pyrrolidinylsulfonyl)phenyl]propa- namide

NP16-XL- 023 3-(5-acetoxylphenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 024 3-(6-acetoxylphenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 025 3-(4-acetoxylphenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 026 3-(6-bromophenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 027 3-(4-methylphenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 028 3-(6-methylphenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 029 3-(4-bromophenyl)-N-[4-(1,2,3,4- tetrahydroquinoline-1-sulfonyl)phenyl]acrylamide

NP16-XL- 030 3-phenyl-N-[4-(1,2,3,4- tetrahydroquinoline-1-sulfonyl)phenyl]acrylamide

NP16-XL- 031 3-(4-bromophenyl)-N-[4-(3,4- dihydro-1H-isoquinoline-2-sulfonyl)phenyl]acrylamide

NP16-XL- 032 3-phenyl-N-[4-(3,4-dihydro-1H- isoquinoline-2-sulfonyl)phenyl]acrylamide

NP16-XL- 035 3-(4-phenylphenyl)-N-[4-(1- indole-1-sulfonyl)phenyl]acrylamide

NP16-XL- 036 3-phenyl-N-{[4-(N-phenyl-3- phenylprop-2-enamido)-sulfonyl]phenyl}-acrylamide

NP16-XL- 037 3-phenyl-N-[(4- phenylsulfamoyl)phenyl]- acrylamide

NP16-XL- 038 3-(6-bromophenyl)-N-[(4- phenylsulfamoyl)phenyl]-acrylamide

NP16-XL- 039 3-(6-bromophenyl)-N-{[4-(N- phenyl-3-6-bromophenylprop-2-enamido)-sulfonyl]phenyl}- acrylamide

NP16-XL- 040 3-(2,6-difluorophenyl)-N-[(4- phenylsulfamoyl)phenyl]-acrylamide

NP16-XL- 041 3-(6-fluorophenyl)-N-[(4- phenylsulfamoyl)phenyl]-acrylamide

NP16-XL- 042 3-(pyridin-3-yl)-N-[(4- phenylsulfamoyl)phenyl]- acrylamide

NP16-XL- 043 3-(6-cyanophenyl)-N-[(4- phenylsulfamoyl)phenyl]-acrylamide

NP16-XL- 044 3-(pyridin-2-yl)-N-[(4- phenylsulfamoyl)phenyl]- acrylamide

NP16-XL- 045 3-(2,6-difluorophenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 046 3-(pyridin-3-yl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 047 3-(6-cyanophenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 048 3-(6-bromophenyl)-N-[(4- phenylamine-carbonyl)phenyl]-acrylamide

NP16-XL- 049 3-(6-cyanophenyl)-N-[(4- phenylamine-carbonyl)phenyl]-acrylamide

NP16-XL- 050 3-(2,6-difluorophenyl)-N-[(4- phenylamine-carbonyl)phenyl]-acrylamide

NP16-XL- 051 3-(6-fluorophenyl)-N-[(4- phenylamine-carbonyl)phenyl]-acrylamide

NP16-XL- 052 3-(6-bromophenyl)-N-[4-(4- methyl-1,4-piperazine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 053 3-(2,6-difluorophenyl)-N-[4-(4- methyl-1,4-piperazine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 054 3-(6-fluorophenyl)-N-[4-(4- methyl-1,4-piperazine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 055 3-(6-cyanophenyl)-N-[4-(4- methyl-1,4-piperazine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 056 3-(pyridin-3-yl)-N-[4-(4-methyl- 1,4-piperazine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 057 4-(5-phenyl-1,3-oxazole)-N-[4- (1-piperidine-1-sulfonyl)phenyl]amide

NP16-XL- 058 3-(2,6-dibromophenyl)-N-[4-(1- piperidine-1-sulfonyl)phenyl]acrylamide

NP16-XL- 059 4-[2-(4-cyanophenyl)-1,5- oxazole]-N-[4-(1-piperidine-1-sulfonyl)phenyl]amide

NP16-XL- 060 4-[2-(thiophen-5-yl)-1,5- oxazole]-N-[4-(1-piperidine-1-sulfonyl)phenyl]amide

One or more compounds of Formula (II) can be combined and/or mixed withone or more of a pharmaceutically acceptable carrier, salt, ester,excipient, vehicle, prodrug, solvent, and diluent to make a composition.

As used herein, the phrase “pharmaceutically acceptable” can meanapproved by a regulatory agency of the Federal or a state government orlisted in the U.S. Pharmacopeia or other generally recognizedpharmacopeia for use in animals and/or in humans.

As used herein, the term “carrier” can refer to a diluent, adjuvant,excipient, and/or vehicle with which the compound and/or antibiotic areadministered. Such pharmaceutical carriers can be sterile liquids, suchas water and oils, including those of petroleum, animal, vegetable orsynthetic origin, such as peanut oil, soybean oil, mineral oil, sesameoil and the like.

As used herein, the phrase “pharmaceutically acceptable salt” can referto derivatives of the compounds defined herein, wherein the parentcompound is modified by making acid or base salts thereof.

The method of treating and/or preventing a microbial infection in asubject can include, but is not limited to, administering to a subjectan effective amount of one or more compounds of Formula (I).

As used herein, the terms “treatment” or “treating” can refer toarresting or inhibiting, or attempting to arrest or inhibit, thedevelopment or progression of an infection and/or causing, or attemptingto cause, the reduction, suppression, regression, or remission of aninfection and/or a symptom thereof. As would be understood by thoseskilled in the art, various clinical and scientific methodologies andassays may be used to assess the development or progression of aninfection, and similarly, various clinical and scientific methodologiesand assays may be used to assess the reduction, regression, or remissionof an infection or its symptoms. “Treatment” refers to both therapeutictreatment and prophylactic or preventative measures. Those in need oftreatment include those already with the infection as well as thoseprone to have the infection or those in whom the infection is to beprevented. In at least some forms, the infection being treated caninclude, but is not limited to, Staphylococcus aureus infection. Inother forms, the infection being treated is a microbial infection.

The administration can include, but is not limited to: administrationthough oral pathways, which administration includes administration incapsule, tablet, granule, spray, syrup, or other such forms;administration through non-oral pathways, which administration includesadministration as an aqueous suspension, an oily preparation or the likeor as a drip, suppository, salve, ointment or the like; administrationvia injection, subcutaneously, intraperitoneally, intravenously,intramuscularly, intradermally, or the like; as well as administrationtopically; and administration via controlled released formulations,depot formulations, and infusion pump delivery.

For intravenous administration, the compounds can be packaged insolutions of sterile isotonic aqueous buffer to make the composition.When necessary, the composition can also include a solubilizing agent.The composition of the compounds can be supplied either separately ormixed together in unit dosage form, for example, as a dry lyophilizedpowder or concentrated solution in a hermetically sealed container suchas an ampoule or sachette indicating the amount of active agent. If thecompound is to be administered by infusion, it can be dispensed with aninfusion bottle containing sterile pharmaceutical grade water or saline.When the compound is administered by injection, an ampoule of sterilewater or saline can be provided so that the ingredients may be mixedprior to injection.

As used herein, the term “subject” can refer to an animal. Typically,the terms “subject” and “patient” may be used interchangeably herein inreference to a subject. As such, a “subject” can include a human that isbeing treated for a microbial infection as a patient.

As used herein, the term “animal” can refer to a mouse, rat, dog, cat,rabbit, pig, monkey, chimpanzee, and human.

As used herein, the terms “effective amount” and “therapeuticallyeffective amount,” can be used interchangeably, as applied to thecompounds, antibiotics, and pharmaceutical compositions described canmean the quantity necessary to render the desired therapeutic result.For example, an effective amount is a level effective to treat, cure, oralleviate the symptoms of an infection for which the composition and/orantibiotic, or pharmaceutical composition, is/are being administered.Amounts effective for the particular therapeutic goal sought will dependupon a variety of factors including the infection being treated and itsseverity and/or stage of development/progression; the bioavailabilityand activity of the specific compound and/or antibiotic, orpharmaceutical composition, used; the route or method of administrationand introduction site on the subject; the rate of clearance of thespecific composition and other pharmacokinetic properties; the durationof treatment; inoculation regimen; drugs used in combination orcoincident with the specific composition; the age, body weight, sex,diet, physiology and general health of the subject being treated; andlike factors well known to one of skill in the relevant scientific art.Some variation in dosage will necessarily occur depending upon thecondition of the subject being treated, and the physician or otherindividual administering treatment will, in any event, determine theappropriate dosage for an individual patient. Furthermore, thetherapeutic methods described would not only apply to treatment in asubject, but could be applied to cell cultures, organs, tissues, orindividual cells in vivo, ex vivo or in vitro.

The term “hydrocarbyl” as used herein includes reference to a moietyconsisting exclusively of hydrogen and carbon atoms; such a moiety maycomprise an aliphatic and/or an aromatic moiety. The moiety may comprise1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20carbon atoms. Examples of hydrocarbyl groups include C₁₋₆ alkyl (e.g.C₁, C₂, C₃ or C₄ alkyl, for example methyl, ethyl, propyl, isopropyl,n-butyl, sec-butyl or tert-butyl); C₁₋₆ alkyl substituted by aryl (e.g.benzyl) or by cycloalkyl (e.g. cyclopropylmethyl); cycloalkyl (e.g.cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl); aryl (e.g. phenyl,naphthyl or fluorenyl) and the like.

The term “alkyl” as used herein includes reference to a straight orbranched chain alkyl moiety having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. Examples of alkylgroups include “C₁₋₆ alkyl” and “C₂₋₁₀ alkyl”. The term “C₁₋₆ alkyl” asused herein include reference to a straight or branched chain alkylmoiety having 1, 2, 3, 4, 5 or 6 carbon atoms. The term “C₂₋₁₀ alkyl” asused herein include reference to a straight or branched chain alkylmoiety having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. This termincludes reference to groups such as methyl, ethyl, propyl (n-propyl orisopropyl), butyl (n-butyl, sec-butyl or tert-butyl), pentyl, hexyl andthe like. In particular, the alkyl moiety may have 1, 2, 3, 4, 5 or 6carbon atoms.

The terms “alkenyl” and “C₂₋₆ alkenyl” as used herein include referenceto a straight or branched chain alkyl moiety having 2, 3, 4, 5 or 6carbon atoms and having, in addition, at least one double bond, ofeither E or Z stereochemistry where applicable. This term includesreference to groups such as ethenyl, 2-propenyl, 1-butenyl, 2-butenyl,3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1-hexenyl, 2-hexenyl and3-hexenyl and the like.

The terms “alkynyl” and “C₂₋₆ alkynyl” as used herein include referenceto a straight or branched chain alkyl moiety having 2, 3, 4, 5 or 6carbon atoms and having, in addition, at least one triple bond. Thisterm includes reference to groups such as ethynyl, 1-propynyl,2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl,3-pentynyl, 1-hexynyl, 2-hexynyl and 3-hexynyl and the like.

The terms “alkoxy” and “C₁₋₆ alkoxy” as used herein include reference to—O-alkyl, wherein alkyl is straight or branched chain and comprises 1,2, 3, 4, 5 or 6 carbon atoms. In one class of embodiments, alkoxy has 1,2, 3 or 4 carbon atoms. This term includes reference to groups such asmethoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentoxy,hexoxy and the like.

The term “cycloalkyl” as used herein includes reference to an alicyclicmoiety having 3, 4, 5, 6, 7 or 8 carbon atoms. The group may be abridged or polycyclic ring system. More often cycloalkyl groups aremonocyclic. This term includes reference to groups such as cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, norbomyl, bicyclo[2.2.2]octyl andthe like.

The term “aryl” as used herein includes reference to an aromatic ringsystem comprising 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 ring carbonatoms. Aryl is often phenyl but may be a polycyclic ring system, havingtwo or more rings, at least one of which is aromatic. This term includesreference to groups such as phenyl, naphthyl, fluorenyl, azulenyl,indenyl, anthryl and the like.

“Cyclic group” means a ring or ring system, which may be unsaturated orpartially unsaturated but is usually saturated, typically containing 5to 13 ring-forming atoms, for example a 5- or 6-membered ring. The ringor ring system may be substituted with one or more hydrocarbyl groups.Cyclic group includes carbocyclyl and heterocyclyl moeities.

The term “carbocyclyl” as used herein includes reference to a saturated(e.g. cycloalkyl) or unsaturated (e.g. aryl) ring moiety having 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 carbon ring atoms. Inparticular, carbocyclyl includes a 3- to 10-membered ring or ring systemand, in particular, 5- or 6-membered rings, which may be saturated orunsaturated. The ring or ring system may be substituted with one or morehydrocarbyl groups. A carbocyclic moiety is, for example, selected fromcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl,bicyclo[2.2.2]octyl, phenyl, naphthyl, fluorenyl, azulenyl, indenyl,anthryl and the like.

The term “heterocyclyl” as used herein includes reference to a saturated(e.g. heterocycloalkyl) or unsaturated (e.g. heteroaryl) heterocyclicring moiety having from 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or16 ring atoms, at least one of which is selected from nitrogen, oxygen,phosphorus, silicon and sulphur. In particular, heterocyclyl includes a3- to 10-membered ring or ring system and more particularly a 5- or6-membered ring, which may be saturated or unsaturated. The ring or ringsystem may be substituted with one or more hydrocarbyl groups.

A heterocyclic moiety is, for example, selected from oxiranyl, azirinyl,1,2-oxathiolanyl, imidazolyl, thienyl, furyl, tetrahydrofuryl, pyranyl,thiopyranyl, thianthrenyl, isobenzofuranyl, benzofuranyl, chromenyl,2H-pyrrolyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrrolizidinyl,imidazolyl, imidazolidinyl, benzimidazolyl, pyrazolyl, pyrazinyl,pyrazolidinyl, thiazolyl, isothiazolyl, dithiazolyl, oxazolyl,isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, piperidyl, piperazinyl,pyridazinyl, morpholinyl, thiomorpholinyl, especially thiomorpholino,indolizinyl, isoindolyl, 3H-indolyl, indolyl, benzimidazolyl, cumaryl,indazolyl, triazolyl, tetrazolyl, purinyl, 4/V-quinolizinyl,isoquinolyl, quinolyl, tetrahydroquinolyl, tetrahydroisoquinolyl,decahydroquinolyl, octahydroisoquinolyl, benzofuranyl, dibenzofuranyl,benzothiophenyl, dibenzothiophenyl, phthalazinyl, naphthyridinyl,quinoxalyl, quinazolinyl, quinazolinyl, cinnolinyl, pteridinyl,carbazoiyl, β-carbolinyl, phenanthridinyl, acridinyl, perimidinyl,phenanthrolinyl, furazanyl, phenazinyl, phenothiazinyl, phenoxazinyl,chromenyl, isochromanyl, chromanyl and the like.

The term “heterocycloalkyl” as used herein includes reference to asaturated heterocyclic moiety having 3, 4, 5, 6 or 7 ring carbon atomsand 1 , 2, 3, 4 or 5 ring heteroatoms selected from nitrogen, oxygen,phosphorus and sulphur. The group may be a polycyclic ring system butmore often is monocyclic. This term includes reference to groups such asazetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, oxiranyl,pyrazolidinyl, imidazolyl, indolizidinyl, piperazinyl, thiazolidinyl,morpholinyl, thiomorpholinyl, quinolizidinyl and the like. The ring orring system may be substituted with one or more hydrocarbyl groups.

The term “heteroaryl” as used herein includes reference to an aromaticheterocyclic ring system having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or16 ring atoms, at least one of which is selected from nitrogen, oxygenand sulphur. The group may be a polycyclic ring system, having two ormore rings, at least one of which is aromatic, but is more oftenmonocyclic. The ring or ring system may be substituted with one or morehydrocarbyl groups. This term includes reference to groups such aspyrimidinyl, furanyl, benzo[b]thiophenyl, thiophenyl, pyrrolyl,imidazolyl, pyrrolidinyl, pyridinyl, benzo[b]furanyl, pyrazinyl,purinyl, indolyl, benzimidazolyl, quinolinyl, phenothiazinyl, triazinyl,phthalazinyl, 2H-chromenyl, oxazolyl, isoxazolyl, thiazolyl, isoindolyl,indazolyl, purinyl, isoquinolinyl, quinazolinyl, pteridinyl and thelike.

The term “halogen” as used herein includes reference to F, Cl, Br or I.

The expression “halogen containing moiety” as used herein includesreference to a moiety comprising 1 to 30 plural valence atoms selectedfrom carbon, nitrogen, oxygen and sulphur which moiety includes at leastone halogen. The moiety may be hydrocarbyl for example C₁₋₆ alkyl orC₁₋₆ alkoxy, or carbocyclyl for example aryl.

The term “substituted” as used herein in reference to a moiety meansthat one or more, especially up to 5, more especially 1, 2 or 3, of thehydrogen atoms in said moiety are replaced independently of each otherby the corresponding number of the described substituents. The term“optionally substituted” as used herein means substituted orun-substituted. It will, of course, be understood that substituents areonly at positions where they are chemically possible, the person skilledin the art being able to decide (either experimentally or theoretically)without inappropriate effort whether a particular substitution ispossible.

The term “enantiomer” as used herein means one of two stereoisomers thathave mirror images of one another.

The term “racemate” as used herein means a mixture of equal amounts ofenantiomers of a chiral molecule.

The term “diastereomer” as used herein means one of a class ofstereoisomers that are not enantiomers, but that have differentconfigurations at one or more of the equivalent chiral centers. Exampleof diasteromers are epimers that differ in configuration of only onechiral center.

The term “stereoisomer” as used herein means one of a class of isomericmolecules that have the same molecular formula and sequence of bondedatoms, but different three-dimensional orientations of their atoms inspace.

The term “prodrug” as used herein refers to a medication that isadministered as an inactive (or less than fully active) chemicalderivative that is subsequently converted to an active pharmacologicalagent in the body, often through normal metabolic processes.

The term “independently” used herein refers to two or more moieties eachselected from a list of atoms or groups, which means that the moietiesmay be the same or different. The identity of each moiety is thereforeindependent of the identities of the one or more other moieties.

The term “jointly” used herein refers to two or more moieties areidentical selected from a list of atoms or groups. In other words, theidentity of each moiety is therefore dependent of the identities of theone or more other moieties being referred to be “jointly” selected fromthe list of atoms or groups.

EXAMPLES

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes in light thereof willbe suggested to persons skilled in the art and are included within thespirit and purview of this application. In addition, any elements orlimitations of any invention or embodiment thereof disclosed herein canbe combined with any and/or all other elements or limitations(individually or in any combination) or any other invention orembodiment thereof disclosed herein, and all such combinations arecontemplated with the scope of the invention without limitation thereto.

Bacteria, Mice, and Chemical Reagents

The strains of S. aureus and E. coli are listed in Table 2. BALB/c micewere purchased from Charles River Laboratories. S. aureus was propagatedin Terrific broth (TB) or on TB agar (Life Technologies; or in BrainHeart Infusion broth (BHI) or on BHI agar (Oxoid). Unless otherwiseindicated, all experiments were performed with bacteria derived fromlight-protected S. aureus 36-48 h stationary phase cultures, the pointat which pigmentation phenotypes were readily apparent.

TABLE 2 Strains Strains Description Source E. coli Rosetta (DE3) Hoststrain for gene expression Lab source S. aureus RN4220 Intermediatecloning host Lab source COL Laboratory strain Lab source AE052 Clinicalisolate [8] COL-ΔcrtN COL with crtN gene replaced ermC This studycassette AE052-ΔcrtN AE052 with crtN gene replaced with This study ermCcassette USA300 CA-MRSA, USA300 FPR3757, ATCC ATCC BAA-1556

Minimum Inhibitory Concentration (MIC) Tests

MIC was determined by inoculating 5×10⁴ S. aureus cells in 100 μl BHImedium in 96-well plates with a serial dilution of antibiotics. The MICwas defined as the minimum concentration resulting in a cell densityless than 0.05 OD at 620 nm, which corresponded to no visible growth,after incubating for 18 h at 37 ° C.

Evaluation of NP-16 Analogues in Staphyloxanthin Production

The in vitro pigment inhibition studies were performed by S. aureusUSA300 cultured in BHI with or without the presence of inhibitorcompounds at 37° C. and 250 rpm for 36-48 hours. The bacteria werewashed twice with PBS prior to the staphyloxanthin purification withmethanol. The OD of the extracts were monitor at 450nm using DTX880multi-plate reader spectrophotometer (Beckman). The concentration rangetested for the compounds were between 300 nM to 700 nM, and controlgroups were added with equal volume of DMSO.

Cytotoxicity Evaluation of Other NP-16 Analogues in Raw 264.7 Cells

The cytotoxicity of NP-16 and some of it analogues in Raw 264.7 cellswas also evaluated by MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assayaccording to manufacturer's instructions. A toxic control (1%) SDS wasincluded to ensure the MTT assay was working properly. The highestconcentration of NP-16 analogues used was 500 μM due to solubilitylimitations. SigmaPlot 11.0 (SPSS, IL) was used for graph plotting.Experiments were carried out in triplicate and repeated twice.

The cytotoxicity of NP-16 and some of it analogues was tested againstRaw 264.7 cells, and the cell tolerance of each compounds weredocumented in Table 3. Along with the in vitro staphyloxanthinproduction of the NP-16 analogues were being tested, the compounds canexert staphyloxanthin production inhibition. The staphyloxanthin fromthe overnight-cultured bacteria were extracted with methanol andquantified in via spectrophotometry. The results were presented inrelative ratio to NP-16 in both the inhibition ratio as well as the TC50in Table 3 (sample names correspond to those listed in Table 1).

TABLE 3 TC50 to Raw Relative ratio to NP16 based Sample Name 264.7 cellson inhibition ratio NP16 >200 1 NP16-XL-010 >200 1.43 NP16-XL-011 751.31 NP16-XL-012 50 1.16 NP16-XL-013 37.5 0.28 NP16-XL-014 >200 0.74NP16-XL-015 >200 1.18 NP16-XL-016 >200 2.71 NP16-XL-017 >200 0.09NP16-XL-018 >200 2.48 NP16-XL-019 >200 1.7 NP16-XL-020 >200 0.17NP16-XL-021 >200 0.76 NP16-XL-022 >200 −0.08 NP16-XL-023 18.7 1.34NP16-XL-024 170 0.43 NP16-XL-025 170 1.92 NP16-XL-026 >200 2.67NP16-XL-027 >200 0.79 NP16-XL-028 >200 2.31 NP16-XL-029 >200 0.53NP16-XL-030 >200 1.26 NP16-XL-031 37.5 0.23 NP16-XL-032 190 1.17NP16-XL-035 50 −0.38 NP16-XL-036 >200 3.04 NP16-XL-037 >200 3.02NP16-XL-038 >200 3.23 NP16-XL-039 >200 3.23 NP16-XL-040 >200 2.51NP16-XL-041 >200 3.17 NP16-XL-042 >200 1.44 NP16-XL-043 >200 3.23NP16-XL-044 200 2.47 NP16-XL-045 200 0.88 NP16-XL-046 >200 −0.05NP16-XL-047 >200 3.18 NP16-XL-048 >200 0.23 NP16-XL-049 50 0.21NP16-XL-050 100 −0.03 NP16-XL-051 150 −0.16 NP16-XL-052 >200 3.18NP16-XL-053 >200 1.26 NP16-XL-054 >200 2.86 NP16-XL-055 >200 3.19NP16-XL-056 >200 0 NP16-XL-057 0.14 NP16-XL-058 0.16 NP16-XL-059 0.06NP16-XL-060 0.06

crtN Expression, Purification and Enzymatic Assay

CrtN with a histidine-maltose binding protein (MBP) tag wasoverexpressed in E. coli Rosetta (DE3) cells. A 10 ml overnight culturewas transferred into 1 L of LB medium supplemented with 100 μg/mlampicillin. Induction was carried out with 1 mM IPTG for 12 hours at 16°C. at an OD of 0.6 at 600 nm. The cell lysate was loaded onto a Ni-NTAcolumn, and CrtN was eluted using a 75-ml linear gradient of 0-0.4 Mimidazole in 50 mM sodium phosphate buffer, with 400 mM sodium chloride,pH 6.6. The collected fractions were analysed by SDS-PAGE to confirm thepeak for MBP-CrtN. The target peak fractions were concentrated and thebuffer was exchanged to loading buffer without imidazole using a PD-10column (GE Healthcare). The collected solution was treated with TEVprotease at 4° C. overnight. The protein sample was applied to a maltosecolumn, and the flow-through was collected as native CrtN protein. Forenzyme assay, 10 μg of purified CrtN was incubated with 100 μl of4,4′-diapophytoene liposomes (containing 5 nmol of 4,4′-diapophytoene),150 μM FAD and buffer II (20 mM phosphate buffer pH 8.0, 100 mM NaCl) ina total volume of 660 μl at 37° C. for 2 h (standard assay). Thereaction was stopped by adding 1 volume of CHCl₃: MeOH (2:1, v/v).Followed by mixing, the sample was centrifuged at 16,000 g for 10 min.The organic phase was dried for LC/MS analysis.

Isolation of Carotenoids

The substrate (4,4′-diapophytoene) and product (4,4′-diaponeurosporene)were extracted from strains COL-ΔcrtN and COL-ΔcrtOP. Carotenoids wereextracted from cell pellets using 300 ml of methanol per liter ofcultured bacteria pellet until all visible pigments were removed. Aftercentrifugation (4° C. and 8,000 g), colored supernatants were pooled andconcentrated to 50 ml using an EZ-2 Plus centrifugal evaporator (GenevacInc., Gardiner, N.Y., USA). A sample was mixed with 100 ml of EtOAc and200 ml of NaCl (2.5 M). The extract sample in the upper organic phasewas collected, washed with same volume of distilled water, and driedusing the EZ-2 Plus evaporator. Dried samples were ready for silica gelisolation or stored at −70 ° C. prior to analysis. For structuralelucidation, carotenoids were identified using a combination of HPLCretention times, UV-visible absorption spectra, and mass fragmentationspectra. Mass fragmentation spectra were monitored using both negativeand positive ion modes in a mass range of m/z 200-1000 on the Varian1200L LC/MS system equipped with an atmospheric pressure chemicalionization interface.

Hydrogen Peroxide Susceptibility Assay

S. aureus was grown in BHI with or without NP16 (40 μM). After 2 days,bacteria were washed twice in PBS, diluted to a concentration of 1×10⁷CFUs per 100 μl reaction mixture in a 96-well plate. Hydrogen peroxide(H₂O₂) in PBS was added to a 440 mM final concentration, and the platewas incubated for 1 hr at 37° C. with shaking. The reaction was stoppedby the addition of 1,000 U/ml of exogenous catalase (Sigma-Aldrich, St.Louis, Mo.), and bacterial viability was assessed by plating dilutionson BHI agar plates.

Bactericidal Activity of Polymorphonuclear Leukocytes

The killing of S. aureus by human polymorphonuclear leukocytes (PMNs)was determined as previously described [9], with some modifications.Briefly, PMNs (10⁶) were mixed with ˜10⁷ opsonized S. aureus bacteriaMOI=10 in 24-well tissue culture plates. After centrifuged at 380 g for8 min, plates were incubated at 37° C. for up to 1.5 h. PMNs were lysedwith saponin (20 min on ice) and plated on BHIA plates. The percentsurvival was calculated by normalized with time zero. Statistics wereperformed with the Student's t-test (GraphPad Prism).

Murine Model of Intraperitoneal Infection

Six- to eight-week-old female Balb/c mice were injected intravenous(i.v) with 1×10⁷ CFUs of early stationary phase S. aureus USA300 orisogenic S. aureus mutant USA300-ΔcrtN. For the treatment study, micewere randomized into groups at the start of the experiment andadministered, i.p. either 17.25 mg/kg of the selected NP-16 analogues orvehicle (5% DMSO with 5% Tween-80) as a control, twice per day. TheNP16, NP16-XL-026 and NP16-XL-043 were administered via intraperitonealroute at 17.25 mg/kg. The kidney bacterial recovery was compared againstvehicle control 7 days after drug treatment.

With the use of intravenous infection of S. aureus USA300 complementedwith isogenic knockouts of −ΔcrtN, mice euthanized on day 7 postinfection with bacterial counts of kidney being quantified, NP16-XL-026and NP16-XL-043 exhibit similar inhibitory effect as NP-16 (FIG. 4)

For other S. aureus sub-type, eight- to ten-week-old female BALB/c micewere injected intraperitoneally (i.p) with 4×10⁸ CFUs of earlystationary phase S. aureus COL. After 3 d, animals were euthanized, theliver and spleen were isolated, homogenized in PBS, and plated on toobtain viable counts. For the treatment study, mice were randomized intotwo groups at the start of the experiment and administered, i.p., either0.35 mg of NP16 or PBS with 5% Tween-80 as a control, twice per day,starting on d −1 to d 2 (a total of eight doses for each).Intraperitoneal challenge with 4×10⁸ CFUs of early stationary phases S.aureus COL was performed on d 0. The mice were sacrificed on d 3 forenumeration of bacterial CFUs in liver and spleen homogenates.

For the clinical isolate S. aureus strain AE052, all operations weresimilar to those used for the COL strain, except 10⁸ CFUs of earlystationary phase bacteria were used in the infection model, and kidneyswere collected for monitoring bacterial loads. Statistics were performedusing the Student's t-test (GraphPad Prism).

Compound NP16 Reduces Pigment Production

Compound NP16 (structure shown in FIG. 1C) had potent activity againstS. aureus pigment formation in vitro, as shown in FIG. 1A, with IC₅₀values ranging from 100 to 300 nM (FIG. 1B). In the biosynthesis ofstaphyloxanthin, the product of CrtN, 4,4′-diaponeurosporene, is ayellowish compound while products prior to CrtM catalysis are colorless.Thus, NP16 treatment is proposed to target CrtM or CrtN or otherregulators that affect the expression of the crtOPQMN cluster, such assigB or ispA [10]. The MIC of NP16 for USA300 was greater than 500 μM(FIG. 1D).

The functions of the five encoded enzymes were characterized by productanalysis of gene deletion mutants. Firstly, in staphyloxanthinbiosynthesis, two molecules of farnesyl diphosphate are condensedhead-to-head to form dehydrosqualene (4,4′-diapophytoene), catalyzed bythe dehydrosqualene synthase CrtM. Secondly, dehydrosqualene isdehydrogenated by the dehydrosqualene desaturase CrtN to form the yellowintermediate 4,4′-diaponeurosporene. Thirdly, oxidation of the terminalmethyl group of 4,4′-diaponeurosporene is catalyzed by a mixed functionoxidase CrtP, to form 4,4′-diaponeurosporenic acid. Then, glycosyl4,4′-diaponeurosporenoate is formed by esterification of glucose at theC1″ position of 4,4′-diaponeurosporenic acid with CrtQ, aglycosyltransferase involved. Finally, glucose at the C6″ position isesterified with the carboxyl group of 12-methyltetradecanoic acid by theacyltransferase CrtO to yield staphyloxanthin. Staphyloxanthin wasidentified as β-D-glucopyranosyl1-O-(4,4′-diaponeurosporen-4-oate)-6-O-(12-methyltetradecanoate).

Inhibition of CrtN by NP16 Results in H₂O₂ and Neutrophil Killing

To probe the biological activities of CrtN, an isogenic crtN mutant inthe COL strain via allelic replacement was generated. The mutationresulted in loss of yellow pigment. Compound NP16 had no effect on thegrowth of MDCK cells (FIG. 2A). A decrease in pigment production wasfound in S. aureus grown in the presence of this NP16 (FIG. 1A).Blocking S. aureus pigment formation has led to an increase in thesusceptibility of the pathogen to hydrogen peroxide killing. For thenon-pigmented strain RN6390, the susceptibility was similar irrespectiveof NP16 treatment (FIG. 2B). Additionally, as a carotenoid producingstrain (FIG. 2D), COL survived significantly better than RN6390 andNP16-treated COL in human neutrophils (FIG. 2C).

Animal Studies

Using a systemic S. aureus infection model, the enzyme CrtM from S.aureus was identified to be a target for anti-infective therapy, basedon virulence factor neutralization [3]. A similar model was applied todetermine if crtN is also essential for infections in mice. The loss ofstaphyloxanthin reduced invasive disease potential, as mice inoculatedwith the isogenic S. aureus mutant COL-ΔcrtN showed lower bacterialpopulation from the liver and spleen, compared with the 4×10⁸ CFUs ofwild-type S. aureus (by intraperitoneal injection), which led to asustained infection (FIGS. 3a and 3b ). Because the COL strain is a lowvirulence strain, no bacteria were detected in the kidneys from day 1 today 3.

Another highly virulent clinical isolate, AE052, and its isogenic S.aureus mutant lacking the CrtN enzyme were also examined by these tests.Compared to wildtype strain, mutant strain in kidney was cleared by hostafter 72 hours post infection (FIG. 3E).

With the same intraperitoneal challenge used in FIGS. 3a, 3b and 3e ,one group of mice (n=14) was treated with 0.35 mg of NP16 twice per day(days −1, 0, 1 and 2), and a second group (n=12) with a vehicle control.Upon sacrificing the mice at 72 hours, S. aureus COL bacterial counts inthe livers and spleens of mice treated with compound NP16 weresignificantly lower than those of the control group (P<0.01) (FIGS. 3cand 3d ). In the case of AE052 infections, bacterial counts in thekidneys of the mice (n=10 for both groups) treated with NP16 weresignificantly lower than those of the control group (P<0.001), with 6 of10 below the detection threshold, versus only 2 of 10 in the controlgroup (FIG. 3F). This result indicates a 98% decrease in survivingbacteria in the treatment groups infected with COL or AE052.

Discussion

It is identified that NP-16 is an inhibitor for CrtN and can exhibitanti-virulence effect on S. aureus. CrtM and CrtN are key enzymes instaphyloxanthin biosynthesis [11]. While staphyloxanthin plays a majorrole in S. aureus tolerance to host defence, it provides a basis forpotential target for rational drug design for the use against S. aureus.It is proposed that a novel anti-infective drug without directbactericidal properties, only targeting mechanisms that renders thepathogens susceptible to normal host innate immune clearance, isprovided. As there is 30% sequence identity between the human SQS andthe bacterial CrtM, and they share significant structural features. Thepresence of such homologue discouraged the employment of CrtM asdruggable target this is further supported by a study focusing on theimprovement of the specificity of BPH652 against CrtM was publishedrecently [12]. Compared with CrtM, CrtN has no homologous enzyme in thehuman cholesterol biosynthesis pathway, making it an attractive drugtarget. A recently proposed CrtN inhibitor, nafitifine, is a topicallyadministered antifungal compound [13], which has been shown to suppresschemotaxis, chemokinesis, chemiluminescence, and superoxide anionproduction of polymorphonuclear leukocytes at high concentrations [14].The effects of naftifine are not stable in different organ (from noeffect to reduced bacterial load for nearly 4 log) and inconsistencywith CrtN mutant (always reduced bacterial load from 0.2 to 2 log atmost). It is believed that this indicates that CrtN should not be theprimary target of naftifine [6].

ROS are employed by phagocytic cells to eliminate bacteria. They aregenerated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase[15]. The bacterial carotenoids expressed by S. aureus may have aprotective function against these defensive molecules [4, 16]. Evidencesupported that a pigment-deficient S. aureus strain was more sensitiveto oxidants, hydrogen peroxide and singlet oxygen, in vitro, as comparedto a wild-type S. aureus strain [1]. Using intra-bacterial inhibitionassay system, showed that the isogenic crtN mutant, which exhibitedinterrupted carotenoid synthesis, was more sensitive to purified humanneutrophils. This confirmed the importance of CrtN in the intracellularsurvival of S. aureus.

CrtN inhibitors without direct bactericidal properties should possesstheoretical advantages of not exerting a direct selective pressure onthe pathogen or normal flora to develop drug resistance. Our approach,as well as other virulence factor-based concepts [3, 17] for highlyspecific anti-staphylococcal therapy relies mainly on the host normalinnate immune response for pathogen clearance. Such strategies are muchmore ideal for clinical treatment and prophylactic applications withlimited risk of developing drug resistant pathogen unlike the caseobserved with antibiotics.

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication.

All patents, patent applications, provisional applications, andpublications referred to or cited herein are incorporated by referencein their entirety, including all figures and tables, to the extent theyare not inconsistent with the explicit teachings of this specification.

It should be understood that numerous specific details, relationships,and methods are set forth to provide a full understanding of theinvention. One having ordinary skill in the relevant art, however, willreadily recognize that the invention can be practiced without one ormore of the specific details or practiced with other methods, protocols,reagents, cell lines and animals. The present invention is not limitedby the illustrated ordering of acts or events, as some acts may occur indifferent orders and/or concurrently with other acts or events.Furthermore, not all illustrated acts, steps or events are required toimplement a methodology in accordance with the present invention. Manyof the techniques and procedures described, or referenced herein, arewell understood and commonly employed using conventional methodology bythose skilled in the art.

Unless otherwise defined, all terms of art, notations and otherscientific terms or terminology used herein are intended to have themeanings commonly understood by those of skill in the art to which thisinvention pertains. In some cases, terms with commonly understoodmeanings are defined herein for clarity and/or for ready reference, andthe inclusion of such definitions herein should not necessarily beconstrued to represent a substantial difference over what is generallyunderstood in the art. It will be further understood that terms, such asthose defined in commonly used dictionaries, should be interpreted ashaving a meaning that is consistent with their meaning in the context ofthe relevant art and/or as otherwise defined herein.

REFERENCE

The following references are incorporated herein by reference in theirentirety:

-   1. Clauditz, A., et al., Staphyloxanthin plays a role in the fitness    of Staphylococcus aureus and its ability to cope with oxidative    stress. Infect Immun, 2006. 74(8): p. 4950-3.-   2. Pelz, A., et al., Structure and biosynthesis of staphyloxanthin    from Staphylococcus aureus. J Biol Chem, 2005. 280(37): p. 32493-8.-   3. Liu, C. I., et al., A cholesterol biosynthesis inhibitor blocks    Staphylococcus aureus virulence. Science, 2008. 319(5868): p.    1391-4.-   4. Liu, G. Y., et al., Staphylococcus aureus golden pigment impairs    neutrophil killing and promotes virulence through its antioxidant    activity. J Exp Med, 2005. 202(2): p. 209-15.-   5. Raisig, A. and G. Sandmann, 4,4′-diapophytoene desaturase:    catalytic properties of an enzyme from the C(30) carotenoid pathway    of Staphylococcus aureus. J Bacteriol, 1999. 181(19): p. 6184-7.-   6. Chen, F., et al., Small-molecule targeting of a diapophytoene    desaturase inhibits S. aureus virulence. Nat Chem Biol, 2016.-   7. Sakai, K., et al., Search method for inhibitors of    Staphyloxanthin production by methicillin-resistant Staphylococcus    aureus. Biol Pharm Bull, 2012. 35(1): p. 48-53.-   8. Ho, P. L., et al., Community-associated methicillin-resistant    Staphylococcus aureus skin and soft tissue infections in Hong Kong.    Hong Kong Med J, 2009. 15 Suppl 9: p. 9-11.-   9. Kobayashi, S. D., et al., Bacterial pathogens modulate an    apoptosis differentiation program in human neutrophils. Proc Natl    Acad Sci USA, 2003. 100(19): p. 10948-53.-   10. Lan, L., et al., Golden pigment production and virulence gene    expression are affected by metabolisms in Staphylococcus aureus. J    Bacteriol, 2010. 192(12): p. 3068-77.-   11. Ku, B., et al., Preparation, characterization, and optimization    of an in vitro C30 carotenoid pathway. Appl Environ Microbiol, 2005.    71(11): p. 6578-83.-   12. Song, Y., et al., Phosphonosulfonates are potent, selective    inhibitors of dehydrosqualene synthase and staphyloxanthin    biosynthesis in Staphylococcus aureus. J Med Chem, 2009. 52(4): p.    976-88.-   13. Favre, B. and N. S. Ryder, Characterization of squalene    epoxidase activity from the dermatophyte Trichophyton rubrum and its    inhibition by terbinafine and other antimycotic agents. Antimicrob    Agents Chemother, 1996. 40(2): p. 443-7.-   14. Vago, T., et al., Effects of naftifine and terbinafine, two    allylamine antifungal drugs, on selected functions of human    polymorphonuclear leukocytes. Antimicrob Agents Chemother, 1994.    38(11): p. 2605-11.-   15. Fang, F. C., Antimicrobial reactive oxygen and nitrogen species:    concepts and controversies. Nat Rev Microbiol, 2004. 2(10): p.    820-32.-   16. Liu, G. Y., et al., Sword and shield: linked group B    streptococcal beta-hemolysin/cytolysin and carotenoid pigment    function to subvert host phagocyte defense. Proc Natl Acad Sci    USA, 2004. 101(40): p. 14491-6.-   17. Sully, E. K., et al., Selective chemical inhibition of agr    quorum sensing in Staphylococcus aureus promotes host defense with    minimal impact on resistance. PLoS Pathog, 2014. 10(6): p. e1004174.-   18. Crossley, K. B., Staphylococci in human disease. 2nd ed. 2010,    Chichester, West Sussex ; Hoboken, N.J.: Wiley-Blackwell. xii, 623    p., 10 p. of plates.-   19. Blot, S. I., et al., Outcome and attributable mortality in    critically Ill patients with bacteremia involving    methicillin-susceptible and methicillin-resistant Staphylococcus    aureus. Arch Intern Med, 2002. 162(19): p. 2229-35.-   20. Peng Gao, Julian Davies and Richard Yi Tsun Kao,    “Dehydrosqualene desaturase as a novel target for antimicrobial    therapeutics in Staphylococcus aureus”, mBio, 8:e01224-17, (2017)

1. A compound having formula (II):

and derivatives thereof, wherein R1 is selected from:

or any four-, five-, six-, seven-, eight-, nine-, ten-, eleven-, ortwelve-membered heterocyclyl, cycloalkenyl, or cycloalkyl, wherein R3and R4 are independently or jointly selected from the group consistingof H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl comprising methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;(C₂₋₄)alkenyl comprising ethenyl, propenyl, butenyl, where a double bondis optionally located at any position in the alkenyl carbon chain, andany alkenyl conformational isomers thereof; alkynyl; aralkyl; alkaryl;halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino; alkylamino;arylamino; dialkylamino; alkylarylamino; diarylamino; acylamino;hydroxyl; thiol; thioalkyl; alkoxy; alkylthio; alkoxyalkyl; aryloxy;arylalkoxy; acyloxy; nitro; carbamoyl; trifluoromethyl; phenoxy;benzyloxy; phosphonic acid; phosphate ester; sulfonic acid (—SO₃H);sulfonate ester; sulfonamide; alkaryl; arylalkyl; carbamate; amino;alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;alkylthio; heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone(═O); ether (—OR10); and ester (—COOR11 and —OC(═O)R11), or R3 and R4are optionally bonded together to form a four-, five-, or six-memberedheterocyclyl, cycloalkenyl, or cycloalkyl, R5 is selected from the groupconsisting of H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl comprising methyl,ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;(C₂₋₄)alkenyl comprising ethenyl, propenyl, butenyl, where a double bondis optionally located at any position in the alkenyl carbon chain, andany alkenyl conformational isomers thereof; alkynyl; aralkyl; alkaryl;halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino; alkylamino;arylamino; dialkylamino; alkylarylamino; diarylamino; acylamino;hydroxyl; thiol; thioalkyl; alkoxy; alkylthio; alkoxyalkyl; aryloxy;arylalkoxy; acyloxy; nitro; carbamoyl; trifluoromethyl; phenoxy;benzyloxy; phosphonic acid; phosphate ester; sulfonic acid (—SO₃H);sulfonate ester; sulfonamide; alkaryl; arylalkyl; carbamate; amino;alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;alkylthio; heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone(═O); ether (—OR10); and ester (—COOR11 and —OC(═O)R11), and wherein R10and R11 are independently or jointly selected from the group consistingof a (C₁₋₄)alkyl comprising methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl comprising ethenyl,propenyl, butenyl, where a double bond is optionally located at anyposition in the alkenyl carbon chain, and any alkenyl conformationalisomers; and alkynyl, X is selected from N or C, A is single bond ordouble bond, Q is selected from N or C, M is selected from O or C, andwherein R2 is selected from:

wherein R6 and R7 are independently or jointly selected from O orabsent; R8 and R9 are independently or jointly selected from the groupconsisting of H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl comprising methyl,ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;(C₂₋₄)alkenyl comprising ethenyl, propenyl, butenyl, where a double bondis optionally located at any position in the alkenyl carbon chain, andany alkenyl conformational isomers thereof; alkynyl; aralkyl; alkaryl;halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino; alkylamino;arylamino; dialkylamino; alkylarylamino; diarylamino; acylamino;hydroxyl; thiol; thioalkyl; alkoxy; alkylthio; alkoxyalkyl; aryloxy;arylalkoxy; acyloxy; nitro; carbamoyl; trifluoromethyl; phenoxy;benzyloxy; phosphonic acid; phosphate ester; sulfonic acid (—SO₃H);sulfonate ester; sulfonamide; alkaryl; arylalkyl; carbamate; amino;alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;alkylthio; heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone(═O); ether (—OR10); and ester (—COOR11 and —OC(═O)R11), or R8 and R9are optionally bonded together to form a four-, five-, or six-memberedheterocyclyl, cycloalkenyl, or cycloalkyl. and wherein R10 and R11 areindependently or jointly selected from the group consisting of a(C₁₋₄)alkyl comprising methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl comprising ethenyl,propenyl, butenyl, where a double bond is optionally located at anyposition in the alkenyl carbon chain, and any alkenyl conformationalisomers; and alkynyl, and Z is selected from C or S.
 2. The compound ofclaim 1, wherein the derivatives comprise:


3. A composition comprising the compound according to claim 1, thederivatives thereof, and a pharmaceutical carrier salt, ester,expicient, vehicle, prodrug, solvent, and diluent, or any combinationthereof.
 4. A composition comprising the compound according to claim 2,the derivatives thereof, and a pharmaceutical carrier salt, ester,excipient, vehicle, prodrug, solvent, and diluent, or any combinationthereof.
 5. A method of treating and/or preventing microbial infectionsand/or related diseases or conditions in a subject comprising:administering to said subject an effective amount of one or morecompounds of formula (II):

wherein R1 is selected from:

or any four-, five-, six-, seven-, eight-, nine-, ten-, eleven-, ortwelve-membered heterocyclyl, cycloalkenyl, or cycloalkyl, wherein R3and R4 are independently or jointly selected from the group consistingof H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl comprising methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;(C₂₋₄)alkenyl comprising ethenyl, propenyl, butenyl, where a double bondis optionally located at any position in the alkenyl carbon chain, andany alkenyl conformational isomers thereof; alkynyl; aralkyl; alkaryl;halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino; alkylamino;arylamino; dialkylamino; alkylarylamino; diarylamino; acylamino;hydroxyl; thiol; thioalkyl; alkoxy; alkylthio; alkoxyalkyl; aryloxy;arylalkoxy; acyloxy; nitro; carbamoyl; trifluoromethyl; phenoxy;benzyloxy; phosphonic acid; phosphate ester; sulfonic acid (—SO₃H);sulfonate ester; sulfonamide; alkaryl; arylalkyl; carbamate; amino;alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;alkylthio; heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone(═O); ether (—OR10); and ester (—COOR11 and —OC(═O)R11), or R3 and R4are optionally bonded together to form a four-, five-, or six-memberedheterocyclyl, cycloalkenyl, or cycloalkyl, R5 is selected from the groupconsisting of H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl comprising methyl,ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;(C₂₋₄)alkenyl comprising ethenyl, propenyl, butenyl, where a double bondis optionally located at any position in the alkenyl carbon chain, andany alkenyl conformational isomers thereof; alkynyl; aralkyl; alkaryl;halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino; alkylamino;arylamino; dialkylamino; alkylarylamino; diarylamino; acylamino;hydroxyl; thiol; thioalkyl; alkoxy; alkylthio; alkoxyalkyl; aryloxy;arylalkoxy; acyloxy; nitro; carbamoyl; trifluoromethyl; phenoxy;benzyloxy; phosphonic acid; phosphate ester; sulfonic acid (—SO₃H);sulfonate ester; sulfonamide; alkaryl; arylalkyl; carbamate; amino;alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;alkylthio; heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone(═O); ether (—OR10); and ester (—COOR11 and —OC(═O)R11), and wherein R10and R11 are independently or jointly selected from the group consistingof a (C₁₋₄)alkyl comprising methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl comprising ethenyl,propenyl, butenyl, where a double bond is optionally located at anyposition in the alkenyl carbon chain, and any alkenyl conformationalisomers; and alkynyl, X is selected from N or C, A is single bond ordouble bond, Q is selected from N or C, M is selected from O or C, andwherein R2 is selected from:

wherein R6 and R7 are independently or jointly selected from O orabsent; R8 and R9 are independently or jointly selected from the groupconsisting of H; F; Cl; Br; I; OH; CN; (C₁₋₄)alkyl comprising methyl,ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl;(C₂₋₄)alkenyl comprising ethenyl, propenyl, butenyl, where a double bondis optionally located at any position in the alkenyl carbon chain, andany alkenyl conformational isomers thereof; alkynyl; aralkyl; alkaryl;halogenated alkyl; heteroalkyl; aryl; heterocyclyl; cycloalkyl;cycloalkenyl; cycloalkynyl; hydroxyalkyl; aminoalkyl; amino; alkylamino;arylamino; dialkylamino; alkylarylamino; diarylamino; acylamino;hydroxyl; thiol; thioalkyl; alkoxy; alkylthio; alkoxyalkyl; aryloxy;arylalkoxy; acyloxy; nitro; carbamoyl; trifluoromethyl; phenoxy;benzyloxy; phosphonic acid; phosphate ester; sulfonic acid (—SO₃H);sulfonate ester; sulfonamide; alkaryl; arylalkyl; carbamate; amino;alkylamino; arylamino; dialkylamino; alkylarylamino; diarylamino;alkylthio; heteroalkyl; alkyltriphenylphosphonium; heterocyclyl; ketone(═O); ether (—OR10); and ester (—COOR11 and —OC(═O)R11), or R8 and R9are optionally bonded together to form a four-, five-, or six-memberedheterocyclyl, cycloalkenyl, or cycloalkyl. and wherein R10 and R11 areindependently or jointly selected from the group consisting of a(C₁₋₄)alkyl comprising methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl; (C₂₋₄)alkenyl comprising ethenyl,propenyl, butenyl, where a double bond is optionally located at anyposition in the alkenyl carbon chain, and any alkenyl conformationalisomers; and alkynyl, and Z is selected from C or S.
 6. The method ofclaim 5, wherein the one or more compounds comprise:


7. The method of claim 5, wherein the microbial infections comprisestaphylococcal infection.
 8. The method of claim 5, wherein the methodreduces the production of pigment in Staphylococcus aureus.
 9. Themethod of claim 5, wherein the microbial infections and/or relateddiseases or conditions comprise infections of the skin and soft tissue,bone and joint, surgical wound, indwelling devices, lung and heartvalves.
 10. The method of claim 5, wherein said subject is a mammal. 11.The method of claim 5, wherein said subject is human.